摘要
为探讨降钙素重组表达质粒(pCEP4-CT)对老龄蛋鸡钙代谢的调控效应。试验首先制备和鉴定降钙素真核表达质粒,而后选用70周龄新扬州鸡36只,随机分为4组,每组3个重复,每个重复3只蛋鸡,研究不同日粮钙(3.5 %和2.5 %)水平条件下,经腿肌注射200μg鸡降钙素基因质粒后对老龄蛋鸡血液中钙代谢相关因子、骨骼质量、养分表观代谢率、蛋壳质量和蛋壳微观构造的影响。结果表明,双酶切电泳和测序结果显示pCEP4-CT质粒制备成功;腿肌注射pCEP4-CT重组表达质粒在蛋鸡体内获得了良好表达,1~14 d时表达量最高,随试验期的延长表达效果逐渐减弱;1~14 d时:同等日粮钙水平条件下,pCEP4-CT质粒可显著提高老龄蛋鸡血清甲状旁腺激素(P<0.05),降低血钙、骨钙素和碱性磷酸酶含量(P<0.05),但对降钙素、雌二醇和血磷含量无显著影响;pCEP4-CT质粒可提高老龄蛋鸡骨骼质量,显著提高胫骨指数和骨密度(P<0.05),促进骨钙和骨磷沉积;pCEP4-CT质粒可促进蛋壳钙沉积,对蛋壳质量和超微构造也有明显改善作用。15~28 d时:注射pCEP4-CT质粒对老龄蛋鸡各项指标均无明显影响。综上所述,pCEP4-CT真核表达质粒可促进老龄蛋鸡对钙的利用,利于骨钙沉积,对蛋壳质量和超微结构也有明显改善作用。饲喂低钙(2.5 %)日粮时,注射pCEP4-CT质粒可显著改善老龄蛋鸡对钙的利用率,促进蛋壳形成,并降低骨质疏松症发病风险。
In order to investigate the effects of calcitonin recombination expression plasmid on the calcium metabolism with old layers. First prepared calcitonin eukaryotic expression plasmid (pCEP4-CT) and identified, then chose 36 New Yangzhou layers in 70 weeks old, which randomly divided into 4 groups of 3 repeat, each repeat contained 3 layers. To study the effect of correlation factor of calcium metabolism, skeleton quality, nutrient appearance metabolic rate, eggshell quality and micro-structure with each layers injected 200μg calcitonin gene plasmid under the different level of calcium in diet (3.5 % and 2.5 %). The results showed that, the pCEP4-CT plasmid prepared succeed by the results of dual-enzyme electrophoresis and sequencing; the pCEP4-CT recombination expression vector expressed well in vivo, between 0~14 d the expression level was highest, and the effect tapered with the experimental stage extend; 0~14 d under the same calcium level, pCEP4-CT plasmid could significantly increase the parathyroid hormone and alkaline phosphatase (P<0.05), and significantly lowering blood calcium and osteocalcin levels (P<0.05), but not affect the estradiol and phosphorus content; pCEP4-CT can improve the bone quality in aging layers, significantly enhance the tibia index number and bone density (P<0.05), and promotion of bone calcium and bone phosphorus deposition; pCEP4-CT plasmid can improve thedeposition of calcium in aging layers, improve the eggshell quality and micro-structure. 15 ~ 28 d , pCEP4-CT plasmid injection had no significant impact on each indicator. In summary, pCEP4-CT eukaryotic expression vector can promote calciummetabolism of laying hens , bone calcium deposition benefit, also improve the eggshell quality and micro-structure. Feeding low calcium (2.5 %) diet, pCEP4-CT plasmid could improve the calcium utilization of aging layers, promote eggshell form, and lower the onset risk of osteoporosis.
引文
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